Department of Microbiology, Cornell University, Ithaca, NY, USA.
Mol Microbiol. 2012 Jun;84(5):845-56. doi: 10.1111/j.1365-2958.2012.08061.x. Epub 2012 May 2.
Agrobacterium tumefaciens incites plant tumours that produce nutrients called opines, which are utilized by the bacteria during host colonization. Various opines provide sources of carbon, nitrogen and phosphorous, but virtually nothing was previously known about how A. tumefaciens acquires sulphur during colonization. Some strains encode an operon required for the catabolism of the opine octopine. This operon contains a gene, msh, that is predicted to direct the conversion of S-methylmethionine (SMM) and homocysteine (HCys) to two equivalents of methionine. Purified Msh carried out this reaction, suggesting that SMM could be an intermediate in opine catabolism. Purified octopine synthase (Ocs, normally expressed in plant tumours) utilized SMM and pyruvate to produce a novel opine, designated sulfonopine, whose catabolism by the bacteria would regenerate SMM. Sulfonopine was produced by tobacco and Arabidopsis when colonized by A. tumefaciens and was utilized as sole source of sulphur by A. tumefaciens. Purified Ocs also used 13 other proteogenic and non-proteogenic amino acids as substrates, including three that contain sulphur. Sulfonopine and 11 other opines were tested for induction of octopine catabolic operon and all were able to do so. This is the first study of the acquisition of sulphur, an essential element, by this pathogen.
根癌农杆菌引发植物肿瘤,产生被称为植物碱的营养物质,这些物质在宿主定殖过程中被细菌利用。各种植物碱为碳、氮和磷提供来源,但以前几乎不知道根癌农杆菌在定殖过程中如何获取硫。一些菌株编码一个操纵子,该操纵子需要分解植物碱章鱼碱。该操纵子包含一个基因 msh,该基因预测指导 S-甲基甲硫氨酸 (SMM) 和同型半胱氨酸 (HCys) 转化为两个当量的蛋氨酸。纯化的 Msh 进行了该反应,表明 SMM 可能是植物碱分解代谢的中间产物。纯化的章鱼碱合酶 (Ocs,通常在植物肿瘤中表达) 利用 SMM 和丙酮酸产生一种新型植物碱,命名为磺化胡椒碱,细菌对其分解代谢可再生 SMM。当烟草和拟南芥被根癌农杆菌定殖时,会产生磺化胡椒碱,根癌农杆菌将其用作唯一的硫源。纯化的 Ocs 还使用 13 种其他蛋白质和非蛋白质氨基酸作为底物,包括 3 种含硫氨基酸。测试了磺化胡椒碱和其他 11 种植物碱是否能诱导章鱼碱分解代谢操纵子的表达,结果都能诱导。这是对该病原体获取必需元素硫的首次研究。